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Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with multi-step derivation method

A technology for simultaneous analysis and carbohydrate metabolism, applied in the field of simultaneous analysis of multi-step derivatization of bio-based samples, to achieve the effect of complete mass spectrogram information, good repeatability, easy retrieval and identification

Active Publication Date: 2012-08-01
湖南省妇幼保健院
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Problems solved by technology

[0007] Aiming at the defects of existing analysis methods such as chromatographic column ion exchange method, organic acid extraction method, urease pretreatment method and Fiehn omics analysis method, the purpose of the present invention is to provide a multi-step derivatization method for synchronous analysis of bases, nuclei Methods for Glycosides, Organic Acids, Fatty Acids, Amino Acids and Carbohydrate Metabolites

Method used

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  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with multi-step derivation method
  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with multi-step derivation method
  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with multi-step derivation method

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Embodiment 1

[0037] 1) Sampling: Use test tube or filter paper to collect urine: use ordinary test tube or domestic No. 3 filter paper, cut into 3.0cmX8.0cm size. Morning urine or any number of urine soaked, filtered and dried, put in a special filter paper drying bag for later use.

[0038] 2) Collect the test sample: dry the filter paper biological matrix sample, elution with 1.0mL deionized double distilled water, and collect the eluate after ultrasonic, centrifugation and other physical methods on the biological sample; determine the creatinine in the eluate Content, determine the creatinine concentration, take 200μl of the eluate equivalent to 2.50μmoL / μl creatinine concentration as the test sample for use;

[0039] 3) Oximation treatment: add 50μl of hydroxylamine hydrochloride aqueous solution (containing 25μg / μl of hydroxylamine hydrochloride) to 200μl of the test sample, adjust the pH of the above test sample to 10 with 6 equivalent NaOH solution to make the test sample complete the ke...

Embodiment 2

[0061] Organisms, microorganisms and plants can be detected under physiological and pathological conditions:

[0062] The preparation method and analysis method are the same as in Example 1, except that by changing the bio-based sample to the laurel bud tissue fluid, the same detection effect as in Example 1 can be obtained, see figure 2 .

[0063] figure 2

[0064] The present invention can also detect other bio-based samples, such as microbial culture fluid, other animal and plant body fluids and tissue fluids, etc. The same detection effect as in Example 1 can be obtained, which will not be listed here.

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Abstract

The invention discloses a method for synchronously analyzing a base, a nucleotide, an organic acid, a fatty acid, an amino acid and a saccharide metabolic product with a multi-step derivation method. The method comprises the following steps of: performing a series of physicochemical method technical treatment such as ultrasonic treatment, centrifugation, oximation, urea removing, phosphorus removing, sulfur removing, protein removing, nitrogen blowing, vacuum drying, and multi-step trimethyl silylation derivation on biological substrate samples (such as urea, blood, cerebrospinal fluid, and tissue fluid); and detecting the biological substrate samples by adopting a gas chromatograph-mass spectrum combination technology. Due to the adoption of method, programmed treatment can be performed on the biological substrate samples simultaneously, and over 1,000 kinds of metabolic intermediate and final products of over five kinds of substances can be detected at one time.

Description

Technical field [0001] The invention belongs to the field of multi-step derivatization synchronous analysis methods for bio-based samples, and specifically relates to a multi-step derivatization method that uses test tubes or filter paper sheets as specimen carriers to simultaneously analyze bases, nucleosides, organic acids, fatty acids, amino acids, and Methods of carbohydrate metabolites. Background technique [0002] Urine contains bases, nucleosides, organic acids, fatty acids, amino acids and carbohydrates and other endogenous compounds such as terminal or intermediate metabolites. Most of these small molecular organic compounds have relatively high water solubility and renal clearance rate Its concentration is usually higher than that of serum and other tissues and body fluids, such as blood, CSF (cerebrospinal fluid) and tissue fluids. Among them, urine specimens are the easiest to collect. It is a non-invasive operation technique and is recognized by researchers at home ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 王益超
Owner 湖南省妇幼保健院
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